White blood cells and neutrophils infiltration.

<p>Cellular infiltration into the lung of mice intratracheally instilled with LPS, TNF-α or vehicle. The amount of White blood cells (WBC), Neutrophils (Neut) and Monocytes (Mono) found in BALF was expressed as number of cells per μl at 3 (A) and 24 hours (B) post treatment. The experiment was repeated three times and each point represents the mean ± standard deviation of 9 animals. Data were expressed as folds of induction over vehicle-treated mice (Saline) and statistical differences were tested by One Way ANOVA followed by Dunnet’s post hoc test for group comparisons. Results are reported as mean ± SD and significance attributed when <i>P</i><0.05 (*) or <i>P</i><0.01(**).</p

bIL-8-Luc reporter construct response <i>in vivo</i>.

<p><b>A</b>) Representative images of groups of mice (n = 3 per group) transiently transgenized with bIL-8-Luc or Empty vector DNA and intratracheally instilled with LPS, TNF-α or vehicle (saline solution). Mice were monitored at 3, 5 and 24 hours post stimulation by BLI drawing a region of interest (ROI) over the chest. <b>B</b>) Light intensity quantification of the ROI using the LivingImage software. The experiment was repeated three times and each point represents the mean ± standard deviation of 9 animals. <b>C</b>) Data were expressed as folds of induction over vehicle-treated mice (Saline) and statistical differences were tested by One Way ANOVA followed by Dunnet’s post hoc test for group comparisons. Results are reported as mean ± SD and significance attributed when <i>P</i><0.05 (*) or <i>P</i><0.01(**).</p

<i>In vitro</i> characterization of bIL-8-Luc reporter construct.

<p><b>A</b>) Schematic diagram (not to scale) of the <i>Bos taurus</i> chromosome 6 with the IL-8 gene locus comprising promoter and exons annotated by numbers. The 2 kb IL-8 promoter sequence, comprising the putative TATA box, the transcriptional start site (+1) and the UTR (red underlined) was cloned into the pGL3 basic vector in front of the open reading frame of the Luciferase reporter. <b>B</b>) RT-PCR products of TLRs from 1 to 9 and their corresponding amplicon size. GAPDH was used as an internal control and the amplification performed in the absence of reverse transcriptase (-RT) as a negative control. <b>C</b>) RT-PCR products of TNF-αRI,II and their corresponding amplicon size. GAPDH was used as an internal control and the amplification performed in the absence of reverse transcriptase (-RT) as a negative control. D) IL-8 promoter activation after treatment of bIL-8-Luc transfected LA-4 cells with LPS or TNF-α, along with the untreated control (Untr.). Each experiment was done in quadruplicate, and each point represents the mean ± standard deviation of three experiments. Data were expressed as folds of induction (2,1 and 2,8 for LPS and TNF-α respectively) over vehicle-treated cells and statistical differences were tested by One Way ANOVA followed by Dunnet’s post hoc test for group comparisons. Results are reported as mean ± SD and significance attributed when <i>P</i><0.05 (*) or <i>P</i><0.01(**).</p

Long lasting response of bIL-8-Luc reporter construct.

<p><b>A</b>) Representative images of groups of mice (n = 3 per group) transiently transgenized with bIL-8-Luc or Empty vector DNA and intratracheally instilled with LPS, TNF-α or vehicle (saline solution) at 8, 32, 45 and 60 days post transgenization. Mice were monitored at 3 hours post stimulation by BLI by drawing a region of interest (ROI) over the chest. <b>B</b>) Light intensity quantification of the ROI using the LivingImage software. The experiment was repeated three times and each point represents the mean ± standard deviation of 9 animals. <b>C</b>) Data were expressed as folds of induction over vehicle-treated mice (Saline) and statistical differences were tested by One Way ANOVA followed by Dunnet’s post hoc test for group comparisons. Results are reported as mean ± SD and significance attributed when <i>P</i><0.05 (*) or <i>P</i><0.01(**).</p

MOESM2 of Monitoring inflammation and airway remodeling by fluorescence molecular tomography in a chronic asthma model

Additional file 2: Figure S2 Representative images of histological slides obtained at 11 weeks from the lungs of mice receiving saline (Panel A) or DRA (Panel B) stained with Alcian/PAS, and from the lungs of mice receiving saline (Panel C) or DRA (Panel D) stained with Masson’s Trichrome. Alcian/PAS and Masson’s Trichrome staining were performed as described in "Methods"

MOESM1 of In vivo monitoring of lung inflammation in CFTR-deficient mice

Additional file 1: Figure S1. In vivo bioluminescence imaging. Monitoring bIL-8 activation in WT and CF transiently transgenized mice with bIL-8-Luc plasmid at 3, 4 and 7 days after DNA delivery. Results are reported photons/sec/cm2 as mean ± SEM, n = 6 each group. Statistical differences were tested by one-way ANOVA followed by Dunnett’s t post hoc test for group comparisons. *p < 0.05 and **p < 0.01

Body weight, number of BALs, and baseline as well as post-BALs (respiratory failure) values of PaO₂ and C_dyn in adult rabbits included in the study.

<p>Body weight, number of BALs, and baseline as well as post-BALs (respiratory failure) values of PaO₂ and C_dyn in adult rabbits included in the study.</p

Histological scores of surfactant-depleted adult rabbits.

<p>Histological scores of surfactant-depleted adult rabbits.</p

Dynamic compliance (C_dyn) of surfactant-depleted rabbits managed with non-invasive ventilation.

<p>To the left of the dashed line, the baseline and post-surfactant depletion C_dyn values of all the animals featured in the study are given. To the right of the dashed line, the C_dyn values of the different groups after 180 minutes of management with NIV are shown. Surfactant-treated groups are represented by the box-plots with a grey filling, whereas the groups merely treated with NIV are represented by the white box-plots. The small squares within each box-plot indicate the mean of the group. The whiskers indicate the maximum and minimum values observed for each group. ^# <i>P</i> < 0.01 vs. SNIPPV+SF group. NCPAP: nasal continuous positive airway pressure, SF: surfactant, NIPPV: nasal (non-synchronized) intermittent positive pressure ventilation, and SNIPPV: synchronized NIPPV.</p

Mean PaO₂ values over time of surfactant-depleted rabbits managed with non-invasive ventilation.

<p>Surfactant-depleted adult rabbits were treated with nasal continuous positive pressure ventilation (NCPAP, white diamonds), with NCPAP + intratracheal surfactant (NCPAP+SF, black diamonds), with nasal non-synchronized intermittent positive ventilation (NIPPV, white squares), with NIPPV + intratracheal surfactant (NIPPV+SF, black squares), with Synchronized Intermittent Positive Pressure Ventilation (SNIPPV, white circles), or with SNIPPV + intratracheal surfactant (SNIPPV+SF, black circles). Surfactant-treated animals (black symbols) received a bolus of surfactant before extubation, immediately after the 0 time-point. Values are shown as the mean ± SEM. * <i>P</i> vs. NIPPV+SF group < 0.01; ^# <i>P</i> vs. SNIPPV group < 0.01; ^§ <i>P</i> vs. NCPAP group < 0.01; ^& <i>P</i> vs. NIPPV group < 0.01.</p